Name: GSM7657449
Instrument: Illumina NovaSeq 6000
Strategy: miRNA-Seq
Source: TRANSCRIPTOMIC
Selection: size fractionation
Layout: SINGLE
Construction protocol: Serum EVs were then isolated via precipitation using “ExoQuick™ - Exosome Precipitation Kit” (#EXOQ20A-1, Biozol, Eching, Germany) according to manufacturer's protocol from 500µL aliquots. The pellet containing precipitated EVs was suspended in 100µL of PBS. 50µL of this suspension were subjected to small RNA extraction using of the “SeraMir™ Exosome RNA Amplification Kit” (#RA808A-1, Biozol, Eching, Germany) with an adapted protocol for extracellular RNA isolation14. The isolates were eluted in 30µL elution buffer and miRNA concentration was quantified using the “Qubit™ microRNA Assay Kit” (#Q32880, Invitrogen™, Waltham, MA) according to the manufacturer's protocol. RNA-sequencing libraries were prepared using the TruSeq® RNA seq Library Prep Kit v2 according to the Illumina TruSeq® messenger (mRNA) sequencing protocol. The RNA-seq libraries were sequenced on the Illumina HiSeq 3000/4000 (1 x 50bp). miRNA-sequencing libraries were prepared according to the manufacturer's protocol of the NEXTFLEX® Small RNA-Seq Kit v3 (#NOVA-5132-06, PerkinElmer, Waltham, MA) and sequenced on SP lanes on an Illumina NovaSeq 6000 (1 × 50 bp).